The field of the invention relates to the use of lipid particles for delivering agents to target cells. In particular, the field of the invention relates to secreted extracellular vesicles (EVs) that contain a targeting membrane protein having an engineered glycosylation site. The secreted extracellular vesicles may be utilized to deliver an agent to a target cell, such as a therapeutic agent.
Secreted extracellular vesicles, such as exosomes, are nanometer-scale lipid vesicles that are produced by many cell types and transfer proteins, nucleic acids, and other molecules between cells in the human body, as well as those of other animals. Targeted exosomes have a wide variety of potential therapeutic uses and have already been shown to be effective for delivery of RNA to neural cells and tumor cells in mice. Here, we describe a method for displaying peptide-based targeting ligands on the exterior of exosomes such that ligands are not degraded by endosomal proteases during exosome biogenesis. This method is novel in that it is the first to acknowledge and address the widespread problem of cleavage of targeting ligands from the luminal terminus extra of integral exosome membrane proteins during exosome biogenesis. Therefore this technology is the first robust method for display of targeting ligands on the exterior of exosomes via the expression of engineered proteins that localize to exosomes. This targeting system can be used for engineering exosomes as targeted gene therapy or drug delivery vehicles in vivo, which could be applied to a wide variety of cell types and diseases.